A Simplified Resectioning Technique

A simple and rapid technique is outlined for 20-40 mUm sectioning of epoxy embedded tissue, removal of areas of interest from the thick sections, resectiening of the selected areas for ultrastructural use. Use of the embeddmg mixture desaibed by Kushida in 1971- makes possible thick sectioning without fracturing. Mounting on glass slides with immersion oil for light microscopic observation facilitates later demouutng for isolation of selected areas. Identification of specific areas of interest is aided by an osmium intensifier stain, p-phenylenediamine. The method is useful for both localizing specific tissue areas and for systematic sampling at precise intervals.     

Effect of 1,3-diaminopropane on ornithine decarboxylase enzyme protein in thioacetamide-treated rat liver.

A radioimmunoassay for ornithine decarboxylase was used to study the regulation of this enzyme in rat liver. The antiserum used reacts with ornithine decarboxylase from mouse, human or rat cells. Rat liver ornithine decarboxylase enzyme activity and enzyme protein (as determined by radioimmunoassay) were measured in thioacetamide-treated rats at various times after administration of 1,3-diaminopropane. Enzyme activity declined rapidly after 1,3-diaminopropane treatment as did the amount of enzyme protein, although the disappearance of enzyme activity slightly preceded the loss of immunoreactive protein. The loss of enzyme protein after cycloheximide treatment also occurred rapidly, but was significantly slower than that seen with 1,3-diaminopropane. When 1,3-diaminopropane and cycloheximide were injected simultaneously, the rate of disappearance of enzyme activity and enzyme protein was the same as that seen with cycloheximide alone. These results show that the rapid loss in enzyme activity after 1,3-diaminopropane treatment is primarily due to a loss in enzyme protein and that protein synthesis is needed in order for 1,3-diaminopropane to exert its full effect. A macromolecular inhibitor of ornithine decarboxylase that has been termed antizyme is induced in response to 1,3-diaminopropane, but our results indicate that the loss of enzyme activity is not due to the accumulation of inactive ornithine decarboxylase-antizyme complexes. It is possible that the antizyme enhances the degradation of the enzyme protein. Control experiments demonstrated that the antiserum used would have detected any inactive antizyme-ornithine decarboxylase complexes present in liver since addition of antizyme to ornithine decarboxylase in vitro did not affect the amount of ornithine decarboxylase detected in our radioimmunoassay. Anti-(ornithine decarboxylase) antibodies may be useful in the purification of antizyme since the antizyme-ornithine decarboxylase complex can be immunoprecipitated, and antizyme released from the precipitate with 0.3 M-NaCl.     

Stereochemistry of the decarboxylation of l-ornithine with ornithine decarboxylase from mouse kidney

Using highly purified ornithine decarboxylase isolated from androgen-treated mice, [1R-²H]putrescine was generated by the decarboxylation of l-ornithine in D₂O, and [1S-²H]putrescine was generated from [2-²H]ornithine by carrying out the decarboxylation in H₂O. Chirality of the putrescines was then determined from the 200-MHz ¹H NMR spectra of their bis-camphanamides in the presence of Eu(fod)₃. These results demonstrated that decarboxylation had taken place with retention of configuration.     

Canada's medical schools

A 20-year-old man with a 10-year history of glomerulonephritis presented with a purpuric rash on his legs. A renal biopsy specimen obtained when he was 11 years old had shown mesangial glomerulonephritis; staining 9 years later for IgA had negative results. A second renal biopsy, performed when the rash was present, revealed mesangial glomerulonephritis and mesangial deposits of IgA; biopsies of the involved skin showed leukocytoclastic vasculitis. In this case isolated glomerulonephritis appeared to change to a multisystem illness, with a different immunologic character, through one of several possible pathogenetic mechanisms.     

The cyclic-2,3-diphosphoglycerate fromMethanobacterium thermoautotrophicum is thed enantiomer

The cyclic pyrophosphate obtained fromMethanobacterium thermoautotrophicum was converted tosn-glycerol 3-phosphate by a stereospecific route. This conversion establishes the structure as cyclic-2,3-diphospho-d-glycerate. The same method was used to determine the cellular content of this metabolite under two conditions: batch culture in a medium containing 2 mM inorganic phosphate and continuous culture with 0.1 mM phosphate in the inflowing medium. The values found were 194±13 and 27.5±1.1 mol/g dry weight, respectively. Computer modeling indicated that the pyrophosphate group cannot adopt a staggered conformation.     

Studies of mammalian ornithine decarboxylase using a monoclonal antibody.

A monoclonal antibody of the immunoglobulin M class was produced against mouse kidney ornithine decarboxylase. Screening for the antibody was carried out using alpha-difluoromethyl[5-3H]ornithine-labelled ornithine decarboxylase. The antibody reacted with this antigen and with native ornithine decarboxylase. The antibody attached to Sepharose could be used to form an immunoaffinity column that retained mammalian ornithine decarboxylase. The active enzyme could then be eluted in a highly purified form by 1.0M-sodium thiocyanate. The monoclonal antibody could also be used to precipitate labelled ornithine decarboxylase from homogenates of kidneys from androgen-treated mice given [35S]methionine. Only one band, corresponding to Mr of about 55000, was observed. The extensive labelling of this band is consistent with the rapid turnover of ornithine decarboxylase protein, since this enzyme represents only about 1 part in 10000 of the cytosolic protein.     

Dynamics of the wax bloom of a seasonal Namib Desert tenebrionid,Cauricara phalangium (Coleoptera: Adesmiini)

Summary The dynamics of the extracuticular was bloom in a winter species of tenebrionid beetle, Cauricara phalangium, from the Namib Desert were monitored in the field and under laboratory conditions. The beetles possessed a full complement of the white wax material after adult emergence. The amount of this material on the integument declined towards the end of the season. The wax bloom was regenerated in both the field and laboratory, with high temperatures and low humidities bringing about greatest renewal. End of the season decline appears tobe related to the senescence of these seasonal beetles. Water loss rates differed significantly for individuals collected in May, when fully bloomed, and in August when little or no wax bloom was present. The wax bloom material contributes to the protection of these diurnal beetles against the high temperatures and radiant heat loads in the Namib Desert.     

Niche-Partitioning of Edaphic Microbial Communities in the Namib Desert Gravel Plain Fairy Circles

Endemic to the Namib Desert, Fairy Circles (FCs) are vegetation-free circular patterns surrounded and delineated by grass species. Since first reported the 1970''s, many theories have been proposed to explain their appearance, but none provide a fully satisfactory explanation of their origin(s) and/or causative agent(s). In this study, we have evaluated an early hypothesis stating that edaphic microorganisms could be involved in their formation and/or maintenance. Surface soils (0–5cm) from three different zones (FC center, FC margin and external, grass-covered soils) of five independent FCs were collected in April 2013 in the Namib Desert gravel plains. T-RFLP fingerprinting of the bacterial (16S rRNA gene) and fungal (ITS region) communities, in parallel with two-way crossed ANOSIM, showed that FC communities were significantly different to those of external control vegetated soil and that each FC was also characterized by significantly different communities. Intra-FC communities (margin and centre) presented higher variability than the controls. Together, these results provide clear evidence that edaphic microorganisms are involved in the Namib Desert FC phenomenon. However, we are, as yet, unable to confirm whether bacteria and/or fungi communities are responsible for the appearance and development of FCs or are a general consequence of the presence of the grass-free circles.     

Phylogeography of the marine macroalga Sargassum hemiphyllum (Phaeophyceae,Heterokontophyta) in northwestern Pacific

Sargassum hemiphyllum is commonly found in Japan and Korea, with a variety, var. chinense, that is found distributed in the southern Chinese coast. We previously reported distinct genetic differentiation between the two taxa based on the PCR‐RFLP data of plastid RubiscoL‐S spacer. The present study aims at elucidating the phylogeographic pattern of S. hemiphyllum based on more markers in the nuclear and extranuclear genomes, with a view to reveal the occurrence of hybridization. The two allopatrically distributed taxa were found to be genetically distinct in nuclear ITS2, plastidial Rubisco (Rbc) and mitochondrial TrnW_I (Trn) spacers. Their divergence was postulated to be attributable to the vicariant event which resulted from the isolation of the Sea of Japan during the late Miocene (6.58–11.25 Mya). Divergence within both S. hemiphyllum and the chinense variety was observed based on Trn spacer, while the divergence in S. hemiphyllum was further confirmed in Rbc spacer. This divergence appears to correspond to the separation of the Japanese populations between the Sea of Japan and the Pacific that occurred around 0.92–2.88 Mya (the early Pleistocene). The presence of an ITS2 clone resembling var. chinense sequences in a Japanese population of S. hemiphyllum (JpNS) raises the possibility of the introgression of var. chinense individuals into S. hemiphyllum population. Compared to that between S. hemiphyllum and the chinense variety, hybridization among the Japanese and Korean populations of S. hemiphyllum is highly probable as all these individuals share a pool of nuclear ITS2 sequences, possibly attributable to incomplete concerted evolution of ITS2.